View source for PCR-from Bacteria

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== Purpose ==
This protocol is used to amplify copies of a desired strand of DNA isolated from a bacterial species.

== Materials Needed ==

== Protocol ==
-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 minimolar dNTPs, 2 microliters of 10 minimolar forward primer, 2 microliters of 10 minimolar reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water

-Suspend one colony of the bacteria in the tube by mixing with a pipette

-Set the PCR machine to have an initial denaturation at 94 C for 5 minutes, 30 cycles of 94 C for 10-30 seconds, 45-65 C for 15-60 seconds and 65 C for 50 seconds per kilobase pair of the DNA fragment, then 65 C for 10 minutes for the final extension, and hold at 4-10 C

-Run the PCR machine with the tube inside

== Notes ==

== Papers where this or a similar method has been used ==
<PMIDsummary />

== Related Ontology Terms ==

== References ==
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[[Category:Methods]]